Core Component B, the Signaling and Motility Assays Core (SMA Core) will provide the reagents, personnel and expertise for the study of motility and motility-related signal transduction, in vitro and in vivo, in adenocarcinoma cells and macrophages. The primary role of the core is to provide a sophisticated initial analysis of motile behavior in a given cell, which has been generated through genetic manipulation, transfection or collection under specific circumstances. Real-time microscopic analysis of cell movement will identify which stages of chemotaxis (lamellipod extension, attachment, contraction, retraction) are altered in the cell of interest. Subsequent analysis will measure changes in the activation of relevant signaling pathways, and test the role of specific signaling proteins by specifically perturbing their function in intact cells. The expertise of the Core will be divided into three areas: (a) light microscopy analysis of the subcellular location and activation state of signaling proteins, both proximal (receptor-related) and distal (actin-related); (b) biochemical analysis of the activation state and physical association of these signaling proteins; and (c) analysis of changes in cell behavior that occur in response to mutation, attenuation or amplification of particular signaling proteins. The core will greatly increase the efficiency of the individual programs by eliminating the need for each separate laboratory to master the wide range of assays needed to characterize the signaling networks linking receptor activation to actin reorganization. The core will standardize the assays analyzing the signaling events and changes in motility that occur upon receptor activation and it will coordinate the use of unique biochemical reagents and methodologies developed by individual laboratories for distribution to all of the projects. Four senior investigators with expertise in biochemistry, cell biology, molecular pharmacology and biological imaging will supervise the Core.